Crossley I, Whalley T, Whitaker M
Department of Physiology, University College London, United Kingdom.
Cell Regul. 1991 Feb;2(2):121-33. doi: 10.1091/mbc.2.2.121.
We show that microinjecting guanosine-5'-thiotriphosphate (GTP gamma S) into unfertilized sea urchin eggs generates an intracellular free calcium concentration [( Ca]i) transient apparently identical in magnitude and duration to the calcium transient that activates the egg at fertilization. The GTP gamma S-induced transient is blocked by prior microinjection of the inositol trisphosphate (InsP3) antagonist heparin. GTP gamma S injection also causes stimulation of the egg's Na+/H+ antiporter via protein kinase C, even in the absence of a [Ca]i increase. These data suggest that GTP gamma S acts by stimulating the calcium-independent production of the phosphoinositide messengers InsP3 and diacylglycerol (DAG). However, the fertilization [Ca]i transient is not affected by heparin, nor can the sperm cause calcium-independent stimulation of protein kinase C. It seems that the bulk of InsP3 and DAG production at fertilization is triggered by the [Ca]i transient, not by the sperm itself. GDP beta S, a G-protein antagonist, does not affect the fertilization [Ca]i transient. Our findings do not support the idea that signal transduction at fertilization operates via a G-protein linked directly to a plasma membrane sperm receptor.
我们发现,向未受精的海胆卵显微注射鸟苷 - 5'-硫代三磷酸(GTPγS)会产生细胞内游离钙浓度[Ca]i瞬变,其幅度和持续时间显然与受精时激活卵子的钙瞬变相同。预先显微注射肌醇三磷酸(InsP3)拮抗剂肝素可阻断GTPγS诱导的瞬变。即使在[Ca]i不增加的情况下,注射GTPγS也会通过蛋白激酶C刺激卵子的Na⁺/H⁺反向转运体。这些数据表明,GTPγS通过刺激磷酸肌醇信使InsP3和二酰基甘油(DAG)的钙非依赖性产生而起作用。然而,受精时的[Ca]i瞬变不受肝素影响,精子也不能引起蛋白激酶C的钙非依赖性刺激。受精时大部分InsP3和DAG的产生似乎是由[Ca]i瞬变触发的,而不是由精子本身触发的。G蛋白拮抗剂GDPβS不影响受精时的[Ca]i瞬变。我们的研究结果不支持受精时信号转导通过直接与质膜精子受体相连的G蛋白起作用这一观点。